Dear Flowers, Is there anybody here who has worked or still working with primer cortical culture (from rat embryo, 3-9 DIV)? I try to analyse Glutamate-induced H2O2 production of cortical cells using DCFH-DA, but I do not know, which population from the three is the proper neural population. I also would greatly appreciate if yo would have some pieces of information on DCFH-DA usage. No I expose cortical culture to 5 mM Glutamate for 6 h, then I wash with DMEM (without phenol red). Then I put 1microL/ml 100-500 microM DCFH-DA (in 0.1% EtOH) to the wells. Incubate for 15 min at 37 celsius, and wash 1x with HBSS (without Ca and Mg) and suspend for measurement. Control value is between 4-20 AFU, Glutamate value is between 6-35 AFU (not a big result) If you have any suggestion, please send me ! Best whishes Andras Boros Gedeon Richter Ltd. Budapest, Hungary
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